LIGHT-INDUCIBLE PROTEIN AGGREGATION SYSTEM FOR MODELING PROTEINOPATHIES AND NEURODEGENERATIVE DISORDERS

摘要

A light-inducible intracellular protein aggregation system is described herein, which provides invaluable tools to study the role of protein aggregates in proteinopathies and to screen for novel therapeutic compounds. The system generally comprises a cell expressing an alpha-synuclein polypeptide or another proteopathic polypeptide that self-aggregates under pathogenic conditions, operably linked to a photoactivatable polypeptide. Illumination of the photoactivatable polypeptide with light having a wavelength sufficient for photoactivation triggers irreversible accumulation of intracellular protein aggregates comprising the alpha-synuclein polypeptide or proteopathic polypeptide. The intracellular protein aggregates can be made to accumulate in real-time during the illumination, thereby enabling spatiotemporal control of protein aggregation. In some embodiments, the intracellular protein aggregates may exhibit pathologically-relevant properties of those found in disease-associated proteinopathies, such as irreversibility, auto-perpetuation (seeding) activity, and comprising misfolded proteins rich in beta-sheet conformation.